Zinc Deficiency Promotes Testicular Cell Apoptosis in Mice

Zinc (Zn) plays an important role in spermatogenesis, and carbon tetrachloride (CCl₄) induces testicular oxidative damage and cell death. The objective of the present study was to define the effects of Zn deficiency in combination with CCl₄ treatment on testicular apoptosis and the associated mechanisms. Mice were fed the following diets with three different Zn levels for 6 weeks: normal zinc (ZN) diet (30 mg Zn/kg), zinc-deficient (ZD) diet (2 mg Zn/kg), and adequate zinc (ZA) diet (100 mg Zn/kg). Beginning in the third week, CCl₄ was intraperitoneally injected into half of the mice in each diet group six times over 3 weeks. We found that Zn was distributed in various tissues and organs in normal mice and that the zinc content in the testis of normal mice was high. The Zn-deficient diet reduced the zinc concentration in the testis tissue, and the testicular/body weight ratio significantly decreased. Moreover, the TUNEL results proved that CCl₄ stimulation of mice fed with a zinc-deficient diet caused marked apoptosis of testicular cells. Furthermore, the ROS levels in the testes obviously increased after Zn-deficient mice were stimulated with CCl₄, whereas reduced glutathione (GSH) and glutathione peroxidase (GSH-Px) showed reduced activities. In addition, proteins associated with the apoptosis signaling pathway were detected with ELISA kits. P-p53, cleaved caspase-3, cleaved PRAP, p-Bad, p-JNK, p-ERK, and p-NF-κB p65 increased by varying degrees under zinc deficiency or CCl₄ stimulation. All the data indicated that Zn deficiency significantly enhanced the harm to the testis induced by oxidative stress and damage, while CCl₄ stimulation exacerbated the oxidative damage in testicular cells, leading to apoptosis through the activation of p53, MAPK, and NF-κB.